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Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy (Articolo in rivista)

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Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy (Articolo in rivista) (literal)

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Hou, YX; Helali, S; Zhang, AD; Jaffrezic-Renault, N; Martelet, C; Minic, J; Gorojankina, T; Persuy, MA; Pajot-Augy, E; Salesse, R; Bessueille, F; Samitier, J; Errachid, A; Akimov, V; Reggiani, L; Pennetta, C; Alfinito, E (2006)
Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy
in Biosensors & bioelectronics
(literal)

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Hou, YX; Helali, S; Zhang, AD; Jaffrezic-Renault, N; Martelet, C; Minic, J; Gorojankina, T; Persuy, MA; Pajot-Augy, E; Salesse, R; Bessueille, F; Samitier, J; Errachid, A; Akimov, V; Reggiani, L; Pennetta, C; Alfinito, E (literal)

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Ecole Cent Lyon, CEGELY, F-69134 Ecully, France; Cent China Normal Univ, Coll Chem, Wuhan 430079, Peoples R China; IPEST, Unite Rech Phys Semicond & Capteurs, La Marsa, Tunisia; INRA, NOPA, RCC, F-78352 Jouy En Josas, France; Lab NanoBioEngn, Barcelona 08028, Spain; Univ Lecce, Dipartimento Ingn Innovaz, I-73100 Lecce, Italy; INFM, Natl Nanotechnol Lab, I-73100 Lecce, Italy (literal)

Titolo

Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy (literal)

Abstract

Rhodopsin, the G protein-coupled receptor (GPCR) which mediates the sense of vision, was prepared from calf eyes and used as receptor enriched membrane fraction. In this study it was immobilized onto gold electrode by two different techniques: Langmuir-Blodgett (LP) and a strategy based on a self-assembled multilayer. We demonstrated that Langmuir and LB films of rhodopsin are not stable. Thus, in this study a new protein multilayer was prepared on gold electrode by building up layer-by-layer a self-assembled multilayer. It is composed of a mixed self-assembled monolayer formed by MHDA and biotinyl-PE, followed by a biotin-avidin system which allows binding of biotinylated antibody specific to rhodopsin. The immobilization of rhodopsin in membrane fraction, by the specific antibody bound previously on self-assembled multilayer, was monitored with electrochemical impedance spectroscopy (EIS). In addition, the specificity and sensitivity of this self-assembled multilayer system to the presence of rhodopsin were investigated. No effect was observed when the system was in contact with olfactory receptor 17 in membrane fraction used for control measurements. All these results demonstrate that rhodopsin can be immobilized efficiently, specifically, quantitatively and stably on gold electrode through the self-assembled multilayer. (c) 2005 Elsevier B.V. All rights reserved. (literal)

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